Research On Tissue Culture Of Cinnamomum Migao

Cinnamomum migao H.W.Li is an evergreen tall tree in Lauraceae native.It is a unique medicinal plant in China,up to 20 meters high.Its dried fruit is one of the top ten seedling medicines in Guizhou Province.It is used to treat heart and gastrointestinal diseases,as well as chest tightness and asthma.C.migao can also be used as industrial raw materials to refine aromatic oils.There is a large demand for the fruits of C.migao in the market due to its medicinal value.In addition,the seed germination rate of C.migao is low,the number of young trees in the C.migao forest is small,and the population naturally reproduces slowly.At present,there are related researches on cutting propagation of C.migao that can solve some problems of rapid propagation technology of them.Tissue culture can shorten the plant reproduction cycle and protect endangered species and their genetic stability.At present,there had been no tissue culture results of C.migao.In this paper,by studying the tissue culture of r C.migao,we explore the best way to sterilize the explants in the tissue culture of C.migao,and the selection and concentration formula of plant hormones in the best induction medium,the best proliferation medium,and the best rooting medium,the best seedling refining method and transplanting substrate to establish a sterile rapid propagation system.The main findings are as follows:(1)Best way to disinfect explants.75% alcohol sterilization for 30 s + 0.1% liter of mercury sterilization for 6.5 min is the best disinfection method for C.migao with budding stem collected indoors.75% alcohol sterilization for 30 s+0.1% liter of mercury for 7 min is the best disinfection method for budding stem of C.migao collected in the field.The rate of bacterial infection of the mussels explants grown indoors was significantly lower than that of the mussels explants collected in the field.When the leaves of C.migao were used as explants,the best disinfection method was:75% alcohol sterilization for 30 s+0.1% liter of mercury for 7 minutes.The infection rate,browning rate,and survival rate of the C.migao explants in different months were different.The contamination rate of budding stem of C.migao collected in April was the lowest,and the budding stem of C.migao collected in November had the highest initiation rate and lower pollution rate.It is the best time for tissue culture explants.(2)Adventitious bud induction.6-BA and IBA are the two most suitable plant hormones forinducing adventitious buds of C.migao,both of which play an important role in adventitious buds induction,and the effect of IBA is greater than 6-BA.The adventitious bud induction rate increased with the increase of 6-BA concentration.The IBA concentration increased continuously,and the induction rate increased first and then decreased.The best medium for induction of adventitious buds with budding stem of C.migao as explants is: MS+6-BA 3.0 mg/L+IBA 0.3mg/L+Agar 6.5 g/L+Sucrose 30 g/L,induction rate is 81.24%.(3)Callus induction.6-BA,NAA,and 2,4-D are suitable for inducing callus of C.migao clams.When the callus was induced by the leaves of C.migao,The effect sequence was: 2,4-D>6-BA>NAA.6-BA and 2,4-D are two important factors affecting the induction rate of callus.Low concentration of 2,4-D,medium concentration of 6-BA and NAA are more suitable for induction of callus.The best induction medium was: MS+2,4-D 1.0 mg/L+6-BA 2.0 mg/L+NAA0.3 mg/L+Agar 6.5 g/L + Sucrose 30 g/L,the induction rate was 88%.(4)Proliferation culture.6-BA and NAA can promote the proliferation of adventitious buds of C.migao.NAA is the main influencing factor for adventitious bud proliferation of C.migao,and the proliferation coefficient increases with the increase of NAA concentration.The optimal propagation medium for adventitious buds of C.migao was: MS + 6-BA 0.1 mg/L+NAA 1.0mg/L+Activated Carbon 0.3 g/L+Agar 6.5 g/L+ Sucrose 30 g/L,and the proliferation coefficient was 2.79.(5)IBA,NAA,and AC were used to induce adventitious bud rooting.The order of the effects of the three hormones on root induction was: IBA> AC > NAA.IBA is the most important factor affecting rooting rate.Low concentration of IBA is easier to induce rooting.AC has a positive effect on the rooting of adventitious buds of C.migao.The best rooting medium for C.migao explants is: 1/2MS + NAA 0.5 mg/L + IBA 0.1 mg/L + AC 0.2 g/L + Agar 7 g/L + Sucrose 30 g/L,the rooting rate reached 72.38%.(6)Refining is a process in which the rooted tissue culture seedlings adapt to the natural growth environment from the artificial environment,which helps increase the survival rate of the seedlings.The longer the time of seedling cultivation,the higher the survival rate of transplanting.And the survival rate of tissue culture seedlings exposed to air earlier was higher.After the cultivating seedlings of C.migao rooting were gradually opened for 7 days,the survival rate oftransplanting can reach 93.1% after 30 days.(7)Different seedlings have their own suitable soil environments.The transplanting substrate can provide water and nutrients for the growth of seedlings,and ensure their survival rate and growth.River sand: Perlite=1:1 is the substrate with the highest survival rate of C.migao seedlings,with a survival rate of 90.63%.The survival rate of C.migao seedlings in the substrate of peat soil:river sand=1:1 was the lowest,with a survival rate of 75%.In the substrate of peat soil:vermiculite=1:1,the leaves of C.migao are lighter in color and have average growth conditions,and the survival rate of seedlings is 76.19%.Peat soil: Perlite=1:1 is the second most important substrate for survival: river sand: Perlite=1:1,with a survival rate of 84.13%,and the C.migao seedlings in this substrate have a fast growth rate and excellent growth.It is the most suitable substrate for the transplantation of C.migao seedlings.